专肽生物_定制多肽合成服务_多肽合成定制服务_多肽合成厂家_多肽合成公司

Peptide YY (PYY) 是一种肠道激素，可调节食欲和抑制胰腺分泌。Peptide YY (PYY) 可以通过神经肽 Y 受体 (Neuropeptide Y receptors) 介导其作用。

Peptide YY (PYY) is a gut hormone that regulates appetite and inhibits pancreatic secretion. Peptide YY (PYY) can mediate its effects through the Neuropeptide Y receptors.

肽YY（PYY）是一种36个氨基酸的肽，是神经肽Y受体Y1、Y2、Y5和Y6的厌食肠道激素激动剂，EC50值分别为0.7、0.58、1和0.8 nM，用于抑制福司柯林诱导的cAMP积累。PYY在体内被切割成PYY（3-36）。PYY的释放发生在餐后，与肠道内分泌L细胞的热量摄入成比例，表明这可能是一种饱腹感信号。在人类中，PYY抑制了瘦和肥胖受试者的食物摄入和胃肠道转运。

Peptide YY (PYY) is a 36-amino acid peptide and anorectic gut hormone agonist for the neuropeptide Y receptors Y1, Y2, Y5, and Y6 with EC50 values of 0.7, 0.58, 1, and 0.8 nM, respectively, for supression of forskolin-induced cAMP accumulation. PYY is cleaved in vivo to PYY (3-36) . Release of PYY occurs postprandially in proportion to calorie intake from intestinal enteroendocrine L cells, indicating it may be a satiety signal. In humans, PYY inhibits food intake and gastrointestinal transit in both lean and obese subjects.

Reference:
[1]. Gerald, C., Walker, M.W., Criscione, L., et al. A receptor subtype involved in neuropeptide-Y-induced food intake. Nature 382(6587), 168-171 (1996).
[2]. Mullins, D.E., Guzzi, M., Xia, L., et al. Pharmacological characterization of the cloned neuropeptide Y y6 receptor. Eur. J. Pharmacol. 395(2), 87-93 (2000).
[3]. Holzer, P., Reichmann, F., and Farzi, A. Neuropeptide Y, peptide YY and pancreatic polypeptide in the gut-brain axis. Neuropeptides 46(6), 261-274 (2012).

多肽H2N-Tyr-Pro-Ile-Lys-Pro-Glu-Ala-Pro-Gly-Glu-Asp-Ala-Ser-Pro-Glu-Glu-Leu-Asn-Arg-Tyr-Tyr-Ala-Ser-Leu-Arg-His-Tyr-Leu-Asn-Leu-Val-Thr-Arg-Gln-Arg-Tyr-NH2的合成步骤：

1、合成MBHA树脂：取若干克的MBHA树脂（如初始取代度为0.5mmol/g）和1倍树脂摩尔量的Fmoc-Linker-OH加入到反应器中，加入DMF，搅拌使氨基酸完全溶解。再加入树脂2倍量的DIEPA，搅拌混合均匀。再加入树脂0.95倍量的HBTU，搅拌混合均匀。反应3-4小时后，用DMF洗涤3次。用2倍树脂体积的10%乙酸酐/DMF 进行封端30分钟。然后再用DMF洗涤3次，甲醇洗涤2次，DCM洗涤2次，再用甲醇洗涤2次。真空干燥12小时以上，得到干燥的树脂{Fmoc-Linker-MHBA Resin}，测定取代度。这里测得取代度为 0.3mmol/g。结构如下图：

2、脱Fmoc：取2.42g的上述树脂，用DCM或DMF溶胀20分钟。用DMF洗涤2遍。加3倍树脂体积的20%Pip/DMF溶液，鼓氮气30分钟，然后2倍树脂体积的DMF 洗涤5次。得到 H2N-Linker-MBHA Resin 。（此步骤脱除Fmoc基团，茚三酮检测为蓝色，Pip为哌啶）。结构图如下：

3、缩合：取2.18mmol Fmoc-Tyr(tBu)-OH 氨基酸，加入到上述树脂里，加适当DMF溶解氨基酸，再依次加入4.36mmol DIPEA，2.07mmol HBTU。反应30分钟后，取小样洗涤，茚三酮检测为无色。用2倍树脂体积的DMF 洗涤3次树脂。（洗涤树脂，去掉残留溶剂，为下一步反应做准备）。得到Fmoc-Tyr(tBu)-Linker-MBHA Resin。氨基酸：DIPEA：HBTU：树脂=3：6：2.85：1（摩尔比）。结构图如下：

4、依次循环步骤二、步骤三，依次得到

H2N-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Pro-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Pro-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Lys(Boc)-Pro-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Lys(Boc)-Pro-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Ile-Lys(Boc)-Pro-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Ile-Lys(Boc)-Pro-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Pro-Ile-Lys(Boc)-Pro-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

H2N-Pro-Ile-Lys(Boc)-Pro-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

Fmoc-Tyr(tBu)-Pro-Ile-Lys(Boc)-Pro-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin

以上中间结构，均可在专肽生物多肽计算器-多肽结构计算器中，一键画出。

最后再经过步骤二得到 H2N-Tyr(tBu)-Pro-Ile-Lys(Boc)-Pro-Glu(OtBu)-Ala-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Ser(tBu)-Pro-Glu(OtBu)-Glu(OtBu)-Leu-Asn(Trt)-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ala-Ser(tBu)-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Leu-Asn(Trt)-Leu-Val-Thr(tBu)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin，结构如下：

5、切割：6倍树脂体积的切割液（或每1g树脂加8ml左右的切割液），摇床摇晃 2小时，过滤掉树脂，用冰无水乙醚沉淀滤液，并用冰无水乙醚洗涤沉淀物3次，最后将沉淀物放真空干燥釜中，常温干燥24小试，得到粗品H2N-Tyr-Pro-Ile-Lys-Pro-Glu-Ala-Pro-Gly-Glu-Asp-Ala-Ser-Pro-Glu-Glu-Leu-Asn-Arg-Tyr-Tyr-Ala-Ser-Leu-Arg-His-Tyr-Leu-Asn-Leu-Val-Thr-Arg-Gln-Arg-Tyr-NH2。结构图见产品结构图。

切割液选择：1）TFA：H2O=95%：5%

2）TFA：H2O：TIS=95%：2.5%：2.5%

3）三氟乙酸：茴香硫醚：1，2-乙二硫醇：苯酚：水=87.5%：5%：2.5%：2.5%：2.5%

（前两种适合没有容易氧化的氨基酸，例如Trp、Cys、Met。第三种适合几乎所有的序列。）

6、纯化冻干：使用液相色谱纯化，收集目标峰液体，进行冻干，获得蓬松的粉末状固体多肽。不过这时要取小样复测下纯度 是否目标纯度。

7、最后总结：

杭州专肽生物技术有限公司（ALLPEPTIDE https://www.allpeptide.com）主营定制多肽合成业务，提供各类长肽，短肽，环肽，提供各类修饰肽，如：荧光标记修饰（CY3、CY5、CY5.5、CY7、FAM、FITC、Rhodamine B、TAMRA等），功能基团修饰肽（叠氮、炔基、DBCO、DOTA、NOTA等），同位素标记肽（N15、C13），订书肽（Stapled Peptide）,脂肪酸修饰肽（Pal、Myr、Ste），磷酸化修饰肽（P-Ser、P-Thr、P-Tyr），环肽（酰胺键环肽、一对或者多对二硫键环），生物素标记肽，PEG修饰肽，甲基化修饰肽等。

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